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Article ; Online: Identification of Deubiquitinase Substrates in Xenopus Egg Extract.

Rossio, Valentina / Paulo, Joao A / King, Randall W

Methods in molecular biology (Clifton, N.J.)

2022 Volume 2591, Page(s) 219–236

Abstract: Deubiquitinases (DUBs) antagonize protein ubiquitination by removing ubiquitin from substrates. Identifying the physiological substrates of each DUB is critical for understanding DUB function and the principles that govern the specificity of this class ... ...

Abstract	Deubiquitinases (DUBs) antagonize protein ubiquitination by removing ubiquitin from substrates. Identifying the physiological substrates of each DUB is critical for understanding DUB function and the principles that govern the specificity of this class of enzymes. Since multiple DUBs can act on the same substrate, it can be challenging to identify substrates using inactivating a single enzyme. Here, we outline a method that enables the identification of proteins whose stability depends on DUB activity and an approach to profile DUB specificity in Xenopus egg extract. By coupling broad DUB inhibition with quantitative proteomics, we circumvent DUB redundancy to identify DUB substrates. By adding back recombinant DUBs individually to the extract, we pinpoint DUBs sufficient to counteract proteasomal degradation of these newly identified substrates. We apply this method to Xenopus egg extract but suggest that it can also be adapted to other cell lysates.
MeSH term(s)	Animals ; Xenopus laevis/metabolism ; Ubiquitin/metabolism ; Ubiquitination ; Proteomics ; Deubiquitinating Enzymes/metabolism
Chemical Substances	Ubiquitin ; Deubiquitinating Enzymes (EC 3.4.19.12)
Language	English
Publishing date	2022-11-06
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-2803-4_13
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Substrate identification and specificity profiling of deubiquitylases against endogenously-generated ubiquitin-protein conjugates.

Rossio, Valentina / Paulo, Joao A / Liu, Xinyue / Gygi, Steven P / King, Randall W

bioRxiv : the preprint server for biology

2023

Abstract: Deubiquitylating enzymes (DUBs) remove ubiquitin from proteins thereby regulating their stability or activity. Our understanding of DUB-substrate specificity is limited because DUBs are typically not compared to each other against many physiological ... ...

Abstract	Deubiquitylating enzymes (DUBs) remove ubiquitin from proteins thereby regulating their stability or activity. Our understanding of DUB-substrate specificity is limited because DUBs are typically not compared to each other against many physiological substrates. By broadly inhibiting DUBs in
Language	English
Publishing date	2023-12-20
Publishing country	United States
Document type	Preprint
DOI	10.1101/2023.12.20.572581
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Book ; Online: Broken Trust

Roth, Randall W. / King, Samuel P.

Greed, Mismanagement & Political Manipulation at America's Largest Charitable Trust

2006

Keywords	History of the Americas
Language	0\|e
Size	1 Online-Ressource
Publisher	University of Hawai'i Press
Document type	Book ; Online
Note	English ; Open Access
HBZ-ID	HT021613645
ISBN	9780824848132 ; 0824848136
Database	ZB MED Catalogue: Medicine, Health, Nutrition, Environment, Agriculture

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Article ; Online: Acceptability of injectable pre-exposure prophylaxis among people who inject drugs in three urban U.S. settings.

King, Adrian R / Shah, Saanchi / Randall, Laura A / Frew, Paula M / Spaulding, Anne / Holloway, Ian W

BMC infectious diseases

2022 Volume 22, Issue 1, Page(s) 737

Abstract: Background: Outbreaks of new HIV transmission among people who inject drugs (PWID) are a major public health concern. Oral daily PrEP, has been identified as a critical addition to the biomedical toolkit for this population. However, limited research on ...

Abstract	Background: Outbreaks of new HIV transmission among people who inject drugs (PWID) are a major public health concern. Oral daily PrEP, has been identified as a critical addition to the biomedical toolkit for this population. However, limited research on the acceptability of long-acting injectable PrEP has been conducted with this population. Methods: We conducted a cross sectional multi-site survey with 1127 participants from May 2019-February 2020 to assess the acceptability of novel PrEP regimens. We computed bivariate and multivariable logistic regressions to evaluate correlates of the outcome variable: acceptability of 3-month injectable-PrEP. SAS v.9.4 was used to conduct statistical analysis. Results: Limited knowledge of or use of PrEP, past or present, was evident within the sample. Injection drug use in the past six months was significantly associated with LA injectable PrEP acceptability, with the odds of acceptability being 1.885 (CI: 1.376, 2.582) times greater than those who did not inject drugs. After adjusting for confounders, injection drug use was significantly associated with the outcome, such that the odds of acceptability of LA injectable PrEP were 1.705 (CI: 1.198, 2.427) times greater among PWID compared to those who did not inject drugs (p < 0.03). The results demonstrate acceptability (38.2%) in a durable (3-month) injectable PrEP modality among participants who also identified as PWID. Conclusions: PrEP promotion efforts among PWID to increase access to long-acting injectable PrEP are necessary. Through efforts to increase acceptance and regular use of long-acting injectable PrEP, public health strategies may be able to effectively lessen chances of future HIV outbreaks among PWID.
MeSH term(s)	Anti-HIV Agents/therapeutic use ; Cross-Sectional Studies ; Drug Users ; HIV Infections/drug therapy ; HIV Infections/epidemiology ; HIV Infections/prevention & control ; Humans ; Pre-Exposure Prophylaxis/methods ; Substance Abuse, Intravenous/complications
Chemical Substances	Anti-HIV Agents
Language	English
Publishing date	2022-09-14
Publishing country	England
Document type	Journal Article
ZDB-ID	2041550-3
ISSN	1471-2334 ; 1471-2334
ISSN (online)	1471-2334
ISSN	1471-2334
DOI	10.1186/s12879-022-07572-3
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Article ; Online: Vaccine confidence among people who use drugs: A cross-sectional survey.

Frew, Paula M / Schamel, Jay T / Randall, Laura A / King, Adrian R / Spaulding, Anne C / Wu, Elizabeth / Holloway, Ian W

Human vaccines & immunotherapeutics

2022 Volume 18, Issue 6, Page(s) 2123201

Abstract: Adult immunization coverage remains low in the US, particularly for people who use drugs (PWUD), a population that experiences a disproportionate burden of vaccine-preventable diseases. The extent of and characteristics associated with vaccine confidence ...

Abstract	Adult immunization coverage remains low in the US, particularly for people who use drugs (PWUD), a population that experiences a disproportionate burden of vaccine-preventable diseases. The extent of and characteristics associated with vaccine confidence (VC) held by PWUD is poorly understood. As VC strongly correlates with vaccine uptake, this cross-sectional study identifies mutable factors associated with VC and quantifies its relationship to immunization status within a highly vulnerable, underimmunized population of PWUD. Using a community-engaged research strategy with select partner organizations hosting syringe exchange programs in Atlanta, Los Angeles, and Las Vegas, USA, we surveyed participants ages 18-69 years served by these organizations from 2019 to 2020. Survey measures included sociodemographics, health behavior including immunization receipt, and vaccine confidence in adult vaccinations using a modified Emory Vaccine Confidence Index (EVCI). The findings reflect relatively low VC among the 1,127 recruited participants, with 56% expressing low VC (EVCI 0-12), 35% medium (EVCI 13-20) and 10% high (EVCI 21-24). EVCI varied by city, with lowest confidence in Atlanta and highest in Las Vegas. VC was associated with past receipt of specific vaccines, including hepatitis A, MMR, Tdap, and influenza. VC varied by specific sociodemographic correlates such as housing insecurity (reduced confidence) and receipt of public benefits or disability (increased confidence). This study identified correlates associated with VC based on site and sociodemographic characteristics for this priority population, highlighting the need for specific interventions to raise VC among PWUD, especially among those experiencing housing insecurity and without public benefits.
MeSH term(s)	Adult ; Humans ; Adolescent ; Young Adult ; Middle Aged ; Aged ; Cross-Sectional Studies ; Vaccination ; Influenza Vaccines ; Immunization ; Vaccination Coverage
Chemical Substances	Influenza Vaccines
Language	English
Publishing date	2022-09-28
Publishing country	United States
Document type	Journal Article
ZDB-ID	2664176-8
ISSN	2164-554X ; 2164-5515
ISSN (online)	2164-554X
ISSN	2164-5515
DOI	10.1080/21645515.2022.2123201
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Article ; Online: Violence Victimization, Homelessness, and Severe Mental Illness Among People Who Use Opioids in Three U.S. Cities.

Hong, Chenglin / Hoskin, Jordan / Berteau, Lorree K / Schamel, Jay T / Wu, Elizabeth S C / King, Adrian R / Randall, Laura A / Holloway, Ian W / Frew, Paula M

Journal of interpersonal violence

2023 Volume 38, Issue 19-20, Page(s) 11165–11185

Abstract: This study examined the associations between violence victimization, homelessness, and severe mental illness (SMI) among people who use opioids (PWUOs) in three U.S. cities. We analyzed data from a cross-sectional survey conducted from May 2019 to ... ...

Abstract	This study examined the associations between violence victimization, homelessness, and severe mental illness (SMI) among people who use opioids (PWUOs) in three U.S. cities. We analyzed data from a cross-sectional survey conducted from May 2019 to February 2020 across three study sites: Los Angeles, CA; Las Vegas, NV; and Atlanta, GA. We used multivariable regressions to examine how multiple victimizations and housing situation are associated with SMI. Based on K-6 scale, nearly half (44.2%) were screened positive for SMI. Meanwhile, 69.7% of the participants reported experiencing some kind of violence in their lifetime, and more than half (51.9%) reported experiencing recent violence (in the past 6 months). The most common form of lifetime violence was emotional (59.5%), followed by physical and intimate partner violence (IPV) (56.1 and 34.9%, respectively), and 34.9% of all participants reported experiencing multiple forms of victimization in the past 6 months. Participants who reported homelessness were more likely to report having experienced recent violence victimization (
MeSH term(s)	Humans ; Analgesics, Opioid/therapeutic use ; Cities ; Cross-Sectional Studies ; Violence ; Intimate Partner Violence/psychology ; Crime Victims/psychology ; Mental Disorders/epidemiology ; Ill-Housed Persons
Chemical Substances	Analgesics, Opioid
Language	English
Publishing date	2023-07-18
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2028900-5
ISSN	1552-6518 ; 0886-2605
ISSN (online)	1552-6518
ISSN	0886-2605
DOI	10.1177/08862605231179720
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Article ; Online: Establishing RTS,S/AS01 as a benchmark for comparison to next-generation malaria vaccines in a mouse model.

Locke, Emily / Flores-Garcia, Yevel / Mayer, Bryan T / MacGill, Randall S / Borate, Bhavesh / Salgado-Jimenez, Berenice / Gerber, Monica W / Mathis-Torres, Shamika / Shapiro, Sarah / King, C Richter / Zavala, Fidel

NPJ vaccines

2024 Volume 9, Issue 1, Page(s) 29

Abstract: New strategies are needed to reduce the incidence of malaria, and promising approaches include vaccines targeting the circumsporozoite protein (CSP). To improve upon the malaria vaccine, RTS,S/AS01, it is essential to standardize preclinical assays to ... ...

Abstract	New strategies are needed to reduce the incidence of malaria, and promising approaches include vaccines targeting the circumsporozoite protein (CSP). To improve upon the malaria vaccine, RTS,S/AS01, it is essential to standardize preclinical assays to measure the potency of next-generation vaccines against this benchmark. We focus on RTS,S/AS01-induced antibody responses and functional activity in conjunction with robust statistical analyses. Transgenic Plasmodium berghei sporozoites containing full-length P. falciparum CSP (tgPb-PfCSP) allow two assessments of efficacy: quantitative reduction in liver infection following intravenous challenge, and sterile protection from mosquito bite challenge. Two or three doses of RTS,S/AS01 were given intramuscularly at 3-week intervals, with challenge 2-weeks after the last vaccination. Minimal inter- and intra-assay variability indicates the reproducibility of the methods. Importantly, the range of this model is suitable for screening more potent vaccines. Levels of induced anti-CSP antibody 2A10 equivalency were also associated with activity: 105 μg/mL (95% CI: 68.8, 141) reduced liver infection by 50%, whereas 285 μg/mL (95% CI: 166, 404) is required for 50% sterile protection from mosquito bite challenge. Additionally, the liver burden model was able to differentiate between protected and non-protected human plasma samples from a controlled human malaria infection study, supporting these models' relevance and predictive capability. Comparison in animal models of CSP-based vaccine candidates to RTS,S/AS01 is now possible under well controlled conditions. Assessment of the quality of induced antibodies, likely a determinant of durability of protection in humans, should be possible using these methods.
Language	English
Publishing date	2024-02-10
Publishing country	England
Document type	Journal Article
ISSN	2059-0105
ISSN (online)	2059-0105
DOI	10.1038/s41541-024-00819-x
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Using cognitive load theory to evaluate and improve preparatory materials and study time for the flipped classroom.

Fischer, Krisztina / Sullivan, Amy M / Cohen, Amy P / King, Randall W / Cockrill, Barbara A / Besche, Henrike C

BMC medical education

2023 Volume 23, Issue 1, Page(s) 345

Abstract: Background: Preclinical medical education is content-dense and time-constrained. Flipped classroom approaches promote durable learning, but challenges with unsatisfactory student preparation and high workload remain. Cognitive load theory defines ... ...

Abstract	Background: Preclinical medical education is content-dense and time-constrained. Flipped classroom approaches promote durable learning, but challenges with unsatisfactory student preparation and high workload remain. Cognitive load theory defines instructional design as "efficient" if learners can master the presented concepts without cognitive overload. We created a PReparatory Evaluation Process (PREP) to systematically assess and measure improvement in the cognitive-load efficiency of preparatory materials and impact on study time (time-efficiency). Methods: We conducted this study in a flipped, multidisciplinary course for ~ 170 first year students at Harvard Medical School using a naturalistic post-test design. For each flipped session (n = 97), we assessed cognitive load and preparatory study time by administering a 3-item PREP survey embedded within a short subject-matter quiz students completed before class. Over three years (2017-2019), we evaluated cognitive load- and time- based efficiency to guide iterative revisions of the materials by content experts. The ability of PREP to detect changes to the instructional design (sensitivity) was validated through a manual audit of the materials. Results: The average survey response rate was ≥ 94%. Content expertise was not required to interpret PREP data. Initially students did not necessarily allocate the most study time to the most difficult content. Over time, the iterative changes in instructional design increased the cognitive load- and time-based efficiency of preparatory materials with large effect sizes (p < .01). Furthermore, this increased the overall alignment of cognitive load with study time: students allocated more time to difficult content away from more familiar, less difficult content without increasing workload overall. Conclusions: Cognitive load and time constraints are important parameters to consider when designing curricula. The PREP process is learner-centered, grounded in educational theory, and works independently of content knowledge. It can provide rich and actionable insights into instructional design of flipped classes not captured by traditional satisfaction-based evaluations.
MeSH term(s)	Humans ; Curriculum ; Learning ; Surveys and Questionnaires ; Education, Medical ; Cognition ; Problem-Based Learning
Language	English
Publishing date	2023-05-17
Publishing country	England
Document type	Journal Article
ZDB-ID	2044473-4
ISSN	1472-6920 ; 1472-6920
ISSN (online)	1472-6920
ISSN	1472-6920
DOI	10.1186/s12909-023-04325-x
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Article: Target-agnostic identification of human antibodies to

Amen, Axelle / Yoo, Randy / Fabra-García, Amanda / Bolscher, Judith / Stone, William J R / Bally, Isabelle / Dergan-Dylon, Sebastián / Kucharska, Iga / de Jong, Roos M / de Bruijni, Marloes / Bousema, Teun / King, C Richter / MacGill, Randall S / Sauerwein, Robert W / Julien, Jean-Philippe / Poignard, Pascal / Jore, Matthijs M

bioRxiv : the preprint server for biology

2024

Abstract: Circulating sexual stages of : Impact statement: A naturally acquired human monoclonal antibody recognizes proteins expressed at different stages of ... ...

Abstract	Circulating sexual stages of Impact statement: A naturally acquired human monoclonal antibody recognizes proteins expressed at different stages of the
Language	English
Publishing date	2024-03-13
Publishing country	United States
Document type	Preprint
DOI	10.1101/2023.11.03.565335
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Article ; Online: Identifying Missed Opportunities for Routine Vaccination among People Who Use Drugs.

Frew, Paula M / Schamel, Jay T / Randall, Laura A / King, Adrian R / Holloway, Ian W / Burris, Katherine / Spaulding, Anne C / Hbou Project Team

International journal of environmental research and public health

2021 Volume 18, Issue 4

Abstract: In the US, adult immunization coverage remains low, especially among vulnerable populations, as recent hepatitis A outbreaks have demonstrated. We studied the vaccination history variation among the US adults who use drugs by implementing a community- ... ...

Abstract	In the US, adult immunization coverage remains low, especially among vulnerable populations, as recent hepatitis A outbreaks have demonstrated. We studied the vaccination history variation among the US adults who use drugs by implementing a community-engaged research survey to identify reported immunization coverage, missed opportunities (MO), and places where immunizations might be delivered. Our analysis of a sample of 1127 participants recruited at community syringe exchanges in three cities identified higher overall vaccination receipt in Los Angeles compared to Atlanta or Las Vegas (e.g., HAV receipt 52.2% LA, 42.1% LV, 41.4% Atlanta). Overall, fewer participants reported having received HAV (45.9%), HBV (47.5%), or influenza (47.6%) vaccines than MMR (57.1%) or Td/Tdap (61.1%). Across sites, HAV receipt was higher for participants incarcerated ≥ 5 years (54.2% vs. 43.6% for those incarcerated < 5 years, 49.4% no incarceration history,
MeSH term(s)	Adult ; Humans ; Influenza Vaccines ; Los Angeles/epidemiology ; Pharmaceutical Preparations ; Vaccination ; Vaccination Coverage
Chemical Substances	Influenza Vaccines ; Pharmaceutical Preparations
Language	English
Publishing date	2021-02-04
Publishing country	Switzerland
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2175195-X
ISSN	1660-4601 ; 1661-7827
ISSN (online)	1660-4601
ISSN	1661-7827
DOI	10.3390/ijerph18041447
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